<abstract xmlns="http://www.w3.org/1999/xhtml"><bold>Introduction:</bold><italic>Mycoplasma gallisepticum</italic> is considered the most pathogenic and economically significant avian <italic>Mycoplasma</italic> spp. for the worldwide poultry industry. The aim of this study was to develop a novel and sensitive real-time loop-mediated isothermal amplification (LAMP) assay based on the amplification of its <italic>mgc2</italic> gene sequence for its rapid molecular detection in poultry.<bold>Material and Methods:</bold> Blood samples from 300 broiler and layer chickens were screened using a rapid serum agglutination (RSA) test. A real-time LAMP reaction was conducted with seropositive swab samples at 60°C for 90 min in an ESEQuant tube scanner using 6-carboxyfluorescein as the reporting dye.<bold>Results:</bold> The sensitivity of the developed assay was 10 fg/μL of DNA. The assay was found 100% specific, showing no cross-reactivity with other avian <italic>Mycoplasma</italic> species. The proportion found of the positive samples by the real-time LAMP was 58%. In comparison, the RSA was found to detect 52% of positive cases.<bold>Conclusion:</bold> The <italic>mgc2</italic> real-time LAMP emerged as a more sensitive and accurate method for molecular detection of <italic>M. gallisepticum</italic> than RSA. Robustness and precision give it applicability as a potential field diagnostic tool for <italic>M. gallisepticum</italic> control. The study will be beneficial in reducing economic losses that <italic>M. gallisepticum</italic> inflicts on the poultry industry. This is the first reported development of a real-time LAMP assay based on the amplification of the <italic>mgc2</italic> gene sequence using an ESEQuant tube scanner for galline <italic>M. gallisepticum</italic> detection.</abstract>

Introduction:Mycoplasma gallisepticum is considered the most pathogenic and economically significant avian Mycoplasma spp. for the worldwide poultry industry. The aim of this study was to develop a novel and sensitive real-time loop-mediated isothermal amplification (LAMP) assay based on the amplification of its mgc2 gene sequence for its rapid molecular detection in poultry.Material and Methods: Blood samples from 300 broiler and layer chickens were screened using a rapid serum agglutination (RSA) test. A real-time LAMP reaction was conducted with seropositive swab samples at 60°C for 90 min in an ESEQuant tube scanner using 6-carboxyfluorescein as the reporting dye.Results: The sensitivity of the developed assay was 10 fg/μL of DNA. The assay was found 100% specific, showing no cross-reactivity with other avian Mycoplasma species. The proportion found of the positive samples by the real-time LAMP was 58%. In comparison, the RSA was found to detect 52% of positive cases.Conclusion: The mgc2 real-time LAMP emerged as a more sensitive and accurate method for molecular detection of M. gallisepticum than RSA. Robustness and precision give it applicability as a potential field diagnostic tool for M. gallisepticum control. The study will be beneficial in reducing economic losses that M. gallisepticum inflicts on the poultry industry. This is the first reported development of a real-time LAMP assay based on the amplification of the mgc2 gene sequence using an ESEQuant tube scanner for galline M. gallisepticum detection.

Real-time loop-mediated isothermal amplification (LAMP) of <italic>mgc2</italic> gene of <italic>Mycoplasma gallisepticum</italic>

University of Veterinary and Animal Sciences

Journal of Veterinary Research

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Real-time loop-mediated isothermal amplification (LAMP) of  gene of 

Introduction:Mycoplasma gallisepticum is considered the most pathogenic and economically significant avian Mycoplasma spp. for the worldwide poultry industry....

Real-time loop-mediated isothermal amplification (LAMP) of mgc2 gene of Mycoplasma gallisepticum

Published Online: Dec 27, 2017

Page range: 439 - 444

Received: Jul 07, 2017

Accepted: Nov 13, 2017

DOI: https://doi.org/10.1515/jvetres-2017-0058

Keywords
poultry, real-time LAMP, gene

© 2017 Syed Ehtisham-ul-Haque et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Real-time loop-mediated isothermal amplification (LAMP) of mgc2 gene of Mycoplasma gallisepticum

Published Online: Dec 27, 2017

Page range: 439 - 444

Received: Jul 07, 2017

Accepted: Nov 13, 2017

DOI: https://doi.org/10.1515/jvetres-2017-0058

Keywordspoultry, real-time LAMP, gene

© 2017 Syed Ehtisham-ul-Haque et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Keywords
poultry, real-time LAMP, gene